Production and characterization of high-titer stocks of rev-defective HIV-1.

Important note: Information in this article was accurate in 1996. The state of the art may have changed since the publication date.

Production and characterization of high-titer stocks of rev-defective HIV-1.

Virology. 1996 Mar 15;217(2):602-6. Unique Identifier : AIDSLINE MED/96183384
Riggs NL; Guatelli JC; San Diego Veterans Affairs Medical Center, University of; California at San Diego, La Jolla 92093-0679, USA.

Abstract: High-titer stocks of rev-defective HIV-1 virions have been produced and characterized. To produce these stocks, CEM cells transduced with a murine retroviral vector that expresses Rev were used to support the growth of an HIV-1 genome containing four inactivating mutations in the rev gene. The resulting viral stocks were of high-titer when used to infect CEM cells that expressed Rev, but had no measurable titer when used to infect parental CEM cells; replication-competent virus was not detected. As expected, these rev-defective viruses established early gene expression in parental CEM cells as demonstrated by the accumulation of viral mRNAs and Nef protein. The expression of late genes was impaired as demonstrated by the inability of these viruses to direct the accumulation of singly spliced mRNA or p24 antigen. These defective viruses did not interfere with replication of the wild-type virus, although they were produced at low levels during coinfection experiments. This experimental system may be useful for the study of early events during the HIV-1 life cycle. In addition, this system represents a simple method of gene transfer into CD4-positive cells using replication-defective but transcriptionally active HIV-1.

Keywords: Base Sequence Cells, Cultured Defective Viruses/*GROWTH & DEVELOPMENT/GENETICS Gene Products, rev/*METABOLISM *Genes, rev HIV-1/GROWTH & DEVELOPMENT/*GENETICS Human Molecular Sequence Data RNA, Viral/BIOSYNTHESIS Support, U.S. Gov't, Non-P.H.S. T-Lymphocytes/MICROBIOLOGY JOURNAL ARTICLE
960830
M9681135

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